Molecular genetic analysis in the diagnosis of lymphoma in fine needle aspiration biopsies. II. Lymphomas versus nonlymphoid malignant tumors

The configurations of immunoglobulin genes and T-cell receptor beta chain genes were analyzed by Southern blotting in DNA derived from nonlymphoid malignant tumors and lymphomas. Gene rearrangements were not detected in any of the 35 cases of nonlymphoid malignant tumors. On the contrary, they were shown in all 14 cases of non-Hodgkin's lymphomas, 2 of 3 cases of Hodgkin's disease and 2 cases diagnosed as non-Hodgkin's lymphoma or angioimmunoblastic lymphadenopathy. The differentiation by light microscopy between lymphoma and nonlymphoid malignant tumors was a diagnostic problem in five cases; the molecular genetic analysis of DNA was contributory in all five diagnostically difficult aspirates. By gene rearrangement studies, the diagnosis of lymphoma was confirmed in two cases and nonlymphoid malignant tumors were accurately indicated in aspirates diagnosed finally as rhabdomyosarcoma (one case) and carcinoma (two cases).     

A human histone H2B.1 variant gene, located on chromosome 1, utilizes alternative 3' end processing.

A variant human H2B histone gene (GL105), previously shown to encode a 2300 nt replication independent mRNA, has been cloned. We demonstrate this gene expresses alternative mRNAs regulated differentially during the HeLa S3 cell cycle. The H2B-Gl105 gene encodes both a 500 nt cell cycle dependent mRNA and a 2300 nt constitutively expressed mRNA. The 3' end of the cell cycle regulated mRNA terminates immediately following the region of hyphenated dyad symmetry typical of most histone mRNAs, whereas the constitutively expressed mRNA has a 1798 nt non-translated trailer that contains the same region of hyphenated dyad symmetry but is polyadenylated. The cap site for the H2B-GL105 mRNAs is located 42 nt upstream of the protein coding region. The H2B-GL105 histone gene was localized to chromosome region 1q21-1q23 by chromosomal in situ hybridization and by analysis of rodent-human somatic cell hybrids using an H2B-GL105 specific probe. The H2B-GL105 gene is paired with a functional H2A histone gene and this H2A/H2B gene pair is separated by a bidirectionally transcribed intergenic promoter region containing consensus TATA and CCAAT boxes and an OTF-1 element. These results demonstrate that cell cycle regulated and constitutively expressed histone mRNAs can be encoded by the same gene, and indicate that alternative 3' end processing may be an important mechanism for regulation of histone mRNA. Such control further increases the versatility by which cells can modulate the synthesis of replication-dependent as well as variant histone proteins during the cell cycle and at the onset of differentiation.     

Assignment of genes encoding a unique cytokine (IL12) composed of two unrelated subunits to chromosomes 3 and 5.

IL12 (formerly NKSF or CLMF) is a unique cytokine composed of two unrelated disulfide-linked subunits. The larger 40-kDa subunit (p40) is a member of the cytokine receptor family, and the smaller 35-kDa subunit (p35) is related to IL6 and GCSF. The chromosomal localization of these two subunits was determined by PCR analysis of DNA from rodent-human hybrids. More refined mapping was obtained by PCR analysis of hybrids containing translocation chromosomes and for p40, by analysis of radiation hybrids. The subunits map to different chromosomes: p40 (IL12B) to 5q31-q33 and p35 (IL12A) to 3p12-3q13.2.     

Two human genes encoding zinc finger proteins,ZNF12 (KOX 3) and ZNF 26 (KOX 20), map to chromosomes 7p22-p21 and 12q24.33, respectively

Summary Two members of the human zinc finger Krüppel family, ZNF 12 (KOX 3) and ZNF 26 (KOX 20), have been localized by somatic cell hybrid analysis and in situ chromosomal hybridization. The presence of individual human zinc finger genes in mouse-human hybrid DNAs was correlated with the presence of specific human chromosomes or regions of chromosomes in the corresponding cell hybrids. Analysis of such mouse-human hybrid DNAs allowed the assignment of the ZNF 12 (KOX 3) gene to chromosome region 7p. The ZNF 26 (KOX 20) gene segregated with chromosome region 12q13-qter. The zinc finger genes ZNF 12 (KOX 3) and ZNF 26 (KOX 20) were localized by in situ chromosomal hybridization to human chromosome regions 7p22-21 and 12q24.33, respectively. These genes and the previously mapped ZNF 24 (KOX 17) and ZNF 29 (KOX 26) genes, are found near fragile sites.     

Serological Studies of the T and Tumor Antigens of the Oncogenic Simian Adenoviruses

Tumor-specific antigens and antisera were prepared for eight of the oncogenic simian adenoviruses. Complement-fixation tests revealed three distinct serological subgroups. This grouping was maintained in studies of virus-infected cells (T antigens) although high titered preparations were obtained for only the major subgroup I. The current grouping is as follows: (I) SV1, SV11, SV25, SV33, SV34, SV38; (II) SV20, SV23; (III) SA 7. Antigens from each subgroup were rapidly inactivated at 56 C, and group II and III antigens were also markedly inactivated at 37 C. One of the tumors (SV1) also contained SV40 T antigen, suggesting origin from a simian adenovirus-SV40 "hybrid."     

RADIOAUTOGRAPHIC LOCALIZATION OF THE INCREASED SYNTHESIS OF PHOSPHATIDYLINOSITOL IN RESPONSE TO PANCREOZYMIN OR ACETYLCHOLINE IN GUINEA PIG PANCREAS SLICES

A technique is described for measuring the incorporation of myo-inositol-2-³H into the lipid of various regions of the guinea pig pancreatic acinar cell by radioautography. Stimulation of enzyme secretion with either pancreozymin or acetylcholine was associated with increased graining in both the basophilic cytoplasm and the nonbasophilic cytoplasm. Kinetic studies suggested that the incorporation of myo-inositol-2-³H was stimulated independently in the two regions. Most of the increment in graining due to stimulation with pancreozymin or acetylcholine plus eserine was abolished if the tissue was extracted with 2:1 chloroform-methanol before radioautography. On chromatography of lipid extracts of pancreas, the only lipid showing a detectable increment in radioactivity on stimulation with pancreozymin was phosphatidylinositol. Thus, essentially all of the increment in graining is likely to be due to increased incorporation of tritium into phosphatidylinositol. These studies, coupled with earlier studies employing differential centrifugation, indicate that on stimulation of enzyme secretion there is increased synthesis of phosphatidylinositol in the rough-surfaced endoplasmic reticulum and in the smooth-surfaced Golgi membranes. The significance of these observations is discussed in connection with membrane circulation presumed to occur in the pancreatic acinar cell on stimulation of protein secretion. It is suggested that the increased synthesis of phosphatidylinositol may be concerned with the formation of new endoplasmic reticulum and possibly Golgi membrane to replace that which is presumably converted to membrane of the zymogen granules during intracellular protein transport.     

Chromosomal localization of human genes required for G1 progression in mammalian cells

Specific probes derived from the human genes that complement the mutations of two independent temperature-sensitive (ts) mutants of the BHK-21 hamster cell line were used to determine the chromosomal locations of the loci in the human genome. The ts11 gene, which complements a mutation that blocks progression through the G1 phase of the cell cycle and which has now been identified as the structural gene for asparagine synthetase, is a member of a small gene/pseudogene family with four members. In a rodent-human somatic cell hybrid panel, the ts11 genomic locus from which the genomic probe derives segregates with human chromosome region 7cen----7q35, proximal to the TCR beta locus. In situ hybridization maps this locus more precisely to the q21-31 region of chromosome 7. Two other members of the gene family detected by the ts11 probe segregate concordantly with chromosome region 8pter----8q24 and chromosome region 21pter----21q22. Similar experiments using the same rodent-human hybrid panel conducted with a probe identifying the tsBN51 gene, which also encodes a function necessary for G1 progression, mapped this locus to human chromosome 8, proximal to the large amplification unit encompassing the c-myc gene of Colo320 cells. Chromosomal in situ hybridization of the tsBN51 probe confirmed the localization of this gene to chromosome 8, with the most likely location of the gene being 8q21.